The objective of this research proposal is to characterize the initial membrane events in hormonal regulation of cellular processes and growth in normal and neoplastically transformed mammalian cells in culture using laser Raman spectroscopy and direct catecholamine binding studies. Using the highly differentiated human lung fibroblast WI-38 and its SV-40 transformed counterpart, WI-38 - VA 13- 2RA (VA13), direct catecholamine binding studies will be related to adenyl cyclase activity in identical systems. Catecholamine binding will be examined in the presence of known beta-antagonists and in the presence of other effector molecules such as Ca2 plus ions, EGTA, adenosine and GTP. To determine the effects the plasma membrane has on this hormone- membrane receptor interaction, as well as subsequent membrane changes which occur with catecholamine binding, laser Raman spectroscopy will be used to characterize both the membrane proteins and lipids in purified plasma membrane vesicles from normal and transformed cells. Plasma membranes from WI-38 cells, VA13 cells, as well as from the mouse fibroblast cell line, Balb/c 3T3 and its SV40 transformed counterpart, Balb-SV-3T3 will be studied to determine changes in receptor-catecholamine interaction by binding studies and membrane structural changes by laser Raman spectroscopy. The catecholamine-membrane receptor interaction will also be investigated in model membrane systems by laser Raman spectroscopy, infrared spectroscopy and fluoresence. Isolated receptor macromolecules will be reincorporated into lipid vesicles to characterize the molecular events in the hormone-receptor interaction. Erythrocyte membrane vesicles will also be used as model systems to develop the use of laser Raman spectroscopy as a probe to study membrane structural changes and their relation to membrane function.